PEDV、TGEV、PARV多重RT-PCR检测方法的建立及其应用

Establishment and clinical application of a multiplex reverse transcription-PCR for detection of porcine epidemic diarrhea virus,porcine transmissible gastroenteritis virus and porcine group A rotavirus

建立了一种同时检测猪流行性腹泻病毒(PEDV)、猪传染性胃肠炎病毒(TGEV)和猪A群轮状病毒(PARV)的三重RT-PCR方法,并对其进行特异性与敏感性试验。在实验室检测中,此方法能检测约50 TCID50的混合病毒,能够扩增出3条长度为750 bp(PEDV)、544 bp(TGEV)、275 bp(PARV)特异性片段,而其他病毒无特异性条带。应用该方法对江苏省及周边地区154份临床疑似病毒性腹泻病料进行检测,结果表明:PEDV、TGEV、PARV阳性率分别为53.2%、8.4%、5.2%。PEDV与TGEV混合感染率为5.2%,PEDV与PARV混合感染率为4.5%。该方法的建立对临床上进行这3种疾病混合感染的检测具有重要意义。

A multiplex reverse transcription polymerase chain reaction （mutiplex RT-PCR） assay was developed for simultaneously detection of porcine epidemic diarrhea virus（ PEDV）, porcine transmissible gastroenteritis virus（TGEV） and porcine group A rotavirus（PARV）. The relative sensitivity and specificity of multiplex RT-PCR were evaluated. In the labo- ratory test, the assay was proved to be sensitive that the detection limit of multiplex RT-PCR was a mixture of 50 TCIDs0 vi- rus. Three specific bands of 750 bp（PEDV） , 544 bp （TGEV） and 275 bp （PARV） were amplified. No specific band was amplified from other pathogenic viruses. 154 samples of small intestinal tissue and faeces of piglets with diarrhea from Jiangsu province and surrounding area were tested by the multiplex RT PCR. The results showed that the positive rates of PEDV, TGEV and PARV were 53.2%, 8.4% and 5.2% respectively. Duple infection of PEDV and TGEV was 5.2%.Duple infection of PEDV and PARV was 4. 5%. This method provides an effective way for detection of co- infection of PEDV, TGEV and PARV from clinical sam- pies.